SPARC基因过表达对卵巢癌淋巴结高转移细胞生物学特性的影响

The Effect of SPARC Gene Over-expression on Biological Characteristics of Human Ovarian Carcinoma Cells with Directional High Lymphatic Metastasis

该研究旨在探讨富含半胱氨酸的酸性分泌蛋白基因(secreted protein acidic and rich in cysteine gene,SPARC)过表达对卵巢癌淋巴结高转移细胞(SKOV3-PM4)生物学特性的影响。构建SPARC基因的慢病毒表达载体并转染SKOV3-PM4细胞,Real-time PCR和Western blot验证转染后的表达效率,激光共聚焦免疫荧光进行蛋白的细胞定位,细胞计数法和集落形成实验测定细胞增殖能力,流式细胞仪检测细胞周期,Transwell小室实验测定细胞体外侵袭、迁移能力。实验结果显示,SPARC蛋白存在于核周及胞质;过表达SPARC基因后,SKOV3-PM4细胞增殖受到明显抑制(P<0.05);细胞周期检测结果显示,各期改变无明显差异;体外侵袭、迁移实验结果显示,SKOV3-PM4细胞侵袭、迁移能力显著降低(P<0.05)。实验结果表明,SPARC基因在卵巢癌淋巴结转移中可能发挥抑癌基因的生物学作用。

The aim of this study was to investigate the effect of secreted protein acidic and rich in cysteine gene（SPARC） overexpression on biological characteristics of human ovarian carcinoma cells with directional high lymphatic metastasis（SKOV3-PM4）. SPARC gene coding region was cloned into lentivirus vector. Lentivirus particles were infected into SKOV3-PM4 cells to upregulate the expression of SPARC gene. The expression level of SPARC was detected at both m RNA and protein levels by Real-time PCR and Western blot. The protein distribution was observed by cell immunofluorescence under laser confocal microscopy. The proliferative ability of SKOV3-PM4 cells was detected by cell counting method and colony formation test, and the cell cycle was measured by flow cytometry. The abilities of cell invasion and migration were examined by Transwell assay. The results showed that SPARC was in perinuclear region and the cytoplasm. Compared with the untransfected cells, cell growth curve and cell colony formation showed that the proliferative ability of SKOV3-PM4-SPARC cells was significantly inhibited（P0.05）. There were no significant differences in cell cycle distribution. The invasion and migration capabilities of SKOV3-PM4-SPARC were also significantly decreased（P0.05）. The abilities of proliferation and invasion of epithelial ovarian cancer cells with directional high lymphatic metastasis were inhibited by SPARC gene over-expression. SPARC gene may play an important role in biological function as a tumor suppressor gene.